Newman P J, Sherman L A
Biochim Biophys Acta. 1978 Aug 8;503(2):343-61. doi: 10.1016/0005-2728(78)90193-7.
Fractions enriched in either Photosystem I or Photosystem II activity have been isolated from the blue-green alga, Synechococcus cedrorum after digitonin treatment. Sedimentation of this homogenate on a 10--30% sucrose gradient yielded three green bands: the upper band was enriched in Photosystem II, the lowest band was enriched in Photosystem I, while the middle band contained both activities. Large quantities of both particles were isolated by zonal centrifugation, and the material was then further purified by chromatography on DEAE-cellulose. The resulting Photosystem II particles carried out light-induced electron transport from semicarbizide to ferricyanide of over 2000 mumol/mg Chlorophyll per h (which was sensitive to 3-(3,4-dichlorophenyl)-1, 1-dimethylurea), and was nearly devoid of Photosystem I activity. This particle contains beta-carotene, very little phycocyanin, has a chlorophyll absorption maximum at 675 nm, and a liquid N2 fluorescence maximum at 685 nm. The purest Photosystem II particles have a chlorophyll to cytochrome b-559 ratio of 50 : 1. The Photosystem I particle is highly enriched in P-700, with a chlorophyll to P-700 ratio of 40 : 1. The physical structure of the two Photosystem particles has also been studied by gel electrophoresis and electron microscopy. These results indicate that the size and protein composition of the two particles are distinctly different.
经洋地黄皂苷处理后,已从蓝绿藻聚球藻中分离出富含光系统I或光系统II活性的组分。将该匀浆在10%-30%的蔗糖梯度上进行沉降,得到三条绿色条带:最上面的条带富含光系统II,最下面的条带富含光系统I,而中间的条带同时含有这两种活性。通过区带离心分离出大量的这两种颗粒,然后将材料进一步通过DEAE-纤维素柱层析进行纯化。所得的光系统II颗粒每小时每毫克叶绿素从氨基脲到铁氰化物的光诱导电子传递超过2000 μmol(对3-(3,4-二氯苯基)-1,1-二甲基脲敏感),并且几乎没有光系统I活性。该颗粒含有β-胡萝卜素,很少有藻蓝蛋白,在675 nm处有叶绿素吸收最大值,在685 nm处有液氮荧光最大值。最纯的光系统II颗粒的叶绿素与细胞色素b-559的比例为50:1。光系统I颗粒高度富含P-700,叶绿素与P-700的比例为40:1。还通过凝胶电泳和电子显微镜研究了这两种光系统颗粒的物理结构。这些结果表明这两种颗粒的大小和蛋白质组成明显不同。